Pharmaceutical Topical Compositions

ABSTRACT

The invention provides pharmaceutical stable semisolid topical compositions comprising between 0.2 to 5% of a des-fluoroquinolone compound, and a suitable carrier to manufacture an ointment or a cream.

This application is a Divisional application of U.S. patent applicationSer. No. 13/124,403, filed Jul. 12, 2011, which is the U.S. NationalPhase of PCT/EP2009/063625, filed Oct. 16, 2009. This application claimspriority under 35 U.S.C. §119(a) to European Patent Application No.08166933.5, filed in Europe on Oct. 17, 2008, the entire contents ofwhich are hereby incorporated by reference.

DESCRIPTION

The present invention concerns topical compositions which comprise ades-fluoroquinolone compound.

Despite advances in antimicrobial therapy over the last 20 years, theincidence of infections caused by multi-drug resistant Gram-positiveorganisms, which are major pathogens in primary uncomplicated skin andskin structure infections (impetigo, foliculitis, forunculosis, acne,secondarily-infected traumatic lesions, overinfected dermatoses, andsecondarily-infected burns) has been increasing. Recently, the increaseof community-acquired methicillin-resistant Staphylococcus aureus (MRSA)infections and the emergence of plasmid-mediated mupirocin resistancealso in MRSA have been reported. Ozenoxacin is a novel non-fluorinatedquinolone compound which has shown a high level of activity againstGram-positive organisms, including common quinolone-resistant bacteria.Ozenoxacin, due to its dual target mechanism of action, is activeagainst some resistant mutant strains. Thus, ozenoxacin is a goodantibacterial agent candidate to circumvent the current mechanisms ofresistance to antibiotics because of its great activity againstresistant Gram-positive bacteria.

Ozenoxacin is active against a great number of pathogens, such asPropionibacterium acnes, Staphylococcus aureus, methicillin-susceptibleStaphylococcus aureus (MSSA), methicillin-resistant Staphylococcusaureus (MRSA) including ciprofloxacin-resistant strains,methicillin-susceptible Staphylococcus epidermidis (MSSE),methicillin-resistant Staphylococcus epidermidis (MRSE), Streptococcuspyogenes, Group G Streptococci, penicillin-resistant Streptococcuspneumoniae, Beta-lactamase positive Haemophilus influenzae, non-typeablestrains of Haemophilus influenzae, Beta-lactamase positive Moraxellacatarrhalis, Neisseria meningitides, Legionella pneumophila, Mycoplasmapneumoniae, Legionella pneumophila, Mycobacterium tuberculosis,Streptococcus agalactiae group B, Neisseria gonorrhoeae, Chlamydiatrachomatis, Mycoplasma hominis, Ureaplasma urealyticum Helicobacterpylori, Bacteroides fragilis, Clostridium perfringens, Escherichia coli,quinolone-resistant Escherichia coli, Salmonella spp., Shigella spp.,ciprofloxacin-susceptible Pseudomonas aeruginosa, Clostridium difficile,and Listeria monocytogenes.

Ozenoxacin (I) was firstly disclosed in U.S. Pat. No. 6,335,447, andequivalent patents. Its chemical name is1-cyclopropyl-8-methyl-7-[5-methyl-6-(methylamino)-3-pyridinyl]-4-oxo-1,4-dihydro-3-quinolinecarboxylicacid. Its chemical formula is:

Topical application of antimicrobial agents is a useful tool for therapyof skin and skin structures infections, sexually transmitted diseasesand genital tract infections and some systemic infections susceptible totopical treatment. Topical antimicrobial therapy has several potentialadvantages compared with systemic therapy. Firstly, it can avoid anunnecessary exposure of the gut flora which may exert selection forresistance. Secondly, it is expected that the high local drugconcentration in topical application and the negligible systemicabsorption should overwhelm many mutational resistances. Thirdly,topical applications are less likely than systemic therapy to cause sideeffects. Accordingly, some topical compositions comprising ozenoxacinhave been reported in the art.

JP2002356426A discloses ointments and gels for skin. An ointmentcomprising ozenoxacin 1%, N-methyl-2-pyrrolidone 8%, propylene glycol14.9%, oleic acid 0.9%, diisopropanolamine 2.3%, polyethylene glycol 40020.2%, polyethylene glycol 4000 50.2%, and water 3.2% is reported inExample 2.

JP2003226643A discloses aqueous solutions comprising ozenoxacin,cyclodextrin, and a viscous agent.

EP1731138A1 discloses fine particle dispersion liquid comprisingozenoxacin to be used in the manufacture of pharmaceutical compositions.

WO2007015453A1 discloses lotions comprising ozenoxacin.

JP2007119456A discloses aqueous suspensions containing nanoparticles andsolution granules of ozenoxacin to be used in the manufacture ofpharmaceutical compositions. Ophthalmic solutions are mentionedpreferably.

A combined use of ozenoxacin, magnesium ions, andhydroxypropyl-β-cyclodextrin specially for ophthalmic use is disclosedin Yamakawa, T. et al., Journal of Controlled Release (2003), 86(1),101-103.

Semisolid topical compositions are useful alternatives to liquidcompositions, because of their better manipulation and consequentpatient preferences. However, in spite of the great diversity ofcomponents present in the semisolid compositions disclosed in the art,no quantitative stability studies are available for them. Thus, there isa need of proved stable semisolid topical compositions comprisingozenoxacin as active ingredient, wherein microbiological and therapeuticactivities are warranted because of demonstrated durable and prolongedpharmaceutical stability.

Accordingly, an object of the present invention relates topharmaceutical stable semisolid topical compositions comprising between0.2 to 5% of the composition of ozenoxacin, and a suitable carrier tomanufacture an ointment or a cream. Preferably, the amount of ozenoxacinis from 0.5% to 2%, and more preferably is 1%. In the present inventionall percentages are expressed in weight percent unless otherwisespecified.

According to the present invention, an ointment wherein the suitablecarrier is selected from white wax, white soft paraffin, and mixturesthereof, is provided. White soft paraffin is preferred.

According to the present invention, a cream wherein the suitable carriercomprises a mixture of emulsifiers, surfactants, oil components, lowmelting point waxes, water, water dispersible components, andnon-formaldehyde-donating preservatives, is provided.

Accordingly, the present invention provides a cream comprising

-   -   a) 0.2-5% of ozenoxacin, and    -   b) a suitable carrier comprising:        -   b.1) 15-25% of one or more emulsifiers;        -   b.2) 10-20% of one or more surfactants;        -   b.3) 5-15% of an oil component;        -   b.4) 1-10% of one or more low melting point waxes selected            from fatty acids having 8 to 30 carbon atoms, fatty alcohols            having 8 to 30 carbon atoms, fatty acid esters having 8 to            30 carbon atoms, fatty acid amides having 8 to 30 carbon            atoms, silicone waxes, and mixtures thereof;        -   b.5) water;        -   b.6) 10-20% of one or more water dispersible components            selected from polyethylene glycol 400, hexylene glycol,            propylene glycol, polypropylene glycol-10 methylglucose            ether, ethoxydiglycol, polyethylene glycol-6 caprylic/capric            glyceride, ethylene glycol monobutyl ether, polyethylene            glycol-8 caprylic/capric glycerides,            3-methoxy-3-methyl-1-butanol, dimethyl isosorbide, and            mixtures thereof; and        -   b.7) 0.01-1% of one or more non-formaldehyde-donating            preservatives;        -   wherein the amount of component b.5 is an amount to complete            100 percent by weight of the composition and all percentages            being weight percent and based on the total weight of the            composition.

According to an embodiment, the present invention provides a creamwherein the carrier comprises:

18 to 22% of component b.1;13 to 15% of component b.2;7 to 9% of component b.3;3 to 5% of component b.4;component b.5;13 to 17% of component b.6;0.05 to 0.15% of component b.7;wherein the amount of component b.5 is an amount to complete 100 percentby weight of the composition.

According to a further embodiment, the present invention provides acream wherein the carrier comprises:

20% of component b.1;14% of component b.2;8% of component b.3;4% of component b.4;component b.5;15% of component b.6;0.1% of component b.7;wherein the amount of component b.5 is an amount to complete 100 percentby weight of the composition.

According to a further embodiment, the amount of water in the carrier is30 to 45% by weight.

In the present invention the emulsifiers are selected from ethyleneglycol monostearate, sorbitan tristearate, a mixture of PEG6 stearate,glycol stearate and PEG32 stearate, and hydrogenated lecithin, andmixtures thereof, preferably the emulsifiers are a mixture of PEG6stearate, glycol stearate and PEG32 stearate.

In the present invention the surfactants are selected from sorbitanoleate monoolein/propylene glycol, C₈/C₁₀ fatty acid mono- anddiglycerides from coconut oil, soy lecithin, egg phosphatides, citricacid esters of monoglycerides, lactic acid esters of monoglycerides,diacetyl tartaric acid esters of monoglycerides, succinic acid esters ofmonoglycerides, sucrose fatty acid esters, polyglycolyzed glycerides ofoleic acids, polyglycolyzed glycerides of linoleic acid, polyglycerolesters of fatty acids, including both long chain and medium chain fattyacids, and polyglyceryl esters of mixed fatty acids, and mixturesthereof. Preferably, the surfactants are polyglycolyzed glycerides ofoleic acids.

In the present invention the oil components are selected from Guerbetalcohols based on fatty alcohols containing 8 to 10 carbon atoms, estersof linear C₆₋₂₂ fatty acids with linear or branched C₆₋₂₂ fattyalcohols, esters of branched C₆₋₁₃ carboxylic acids with linear orbranched C₆₋₂₂ fatty alcohols, esters of linear C₆₋₂₂ fatty acids with2-ethyl hexanol, esters of C₃₋₃₈ alkylhydroxycarboxylic acids withlinear or branched C₆₋₂₂ fatty alcohols, esters of linear or branchedfatty acids with polyhydric alcohols or Guerbet alcohols, triglyceridesbased on C₆₋₁₀ fatty acids, liquid mono-, di- and triglyceride mixturesbased on C₆₋₁₈ fatty acids, esters of C₆₋₂₂ fatty alcohols or Guerbetalcohols with benzoic acid, esters of C₂₋₁₂ dicarboxylic acids withlinear or branched alcohols containing 1 to 22 carbon atoms or polyolscontaining 2 to 10 carbon atoms and 2 to 6 hydroxyl groups, vegetableoils, branched primary alcohols, substituted cyclohexanes, linear andbranched C₆₋₂₂ fatty alcohol carbonates, Guerbet carbonates based onfatty alcohols containing 8 to 10 carbon atoms, esters of benzoic acidwith linear or branched C₆₋₂₂ alcohols, linear or branched, symmetricalor nonsymmetrical dialkyl ethers containing 6 to 22 carbon atoms peralkyl group, ring opening products of epoxidized fatty acid esters withpolyols, silicone oils, and aliphatic or naphthenic hydrocarbons, andmixtures thereof. Preferably, the oil components are Guerbet alcoholsbased on fatty alcohols containing 8 to 10 carbon atoms, such as 2-octyldodecanol (Eutanol® G PH).

Because they are primary, branched, and of high molecular weight,Guerbet alcohols have low irritation potential, are liquid to extremelylow temperatures, are low in volatility, are useful as superfattingagents and are good lubricants.

In the present invention the low melting point waxes are selected fromfatty acids having 8 to 30 carbon atoms, fatty alcohols having 8 to 30carbon atoms, fatty acid esters having 8 to 30 carbon atoms, fatty acidamides having 8 to 30 carbon atoms, silicone waxes, and mixturesthereof. Preferably, the low melting point waxes are fatty alcoholshaving 8 to 30 carbon atoms. More preferably, stearyl alcohol isselected from fatty alcohols.

On account of its consistency giving characteristics, stearyl alcoholacts as a convenient viscosity regulator.

In the present invention the water dispersible components are selectedfrom polyethylene glycol 400, hexylene glycol, propylene glycol,polypropylene glycol-10 methylglucose ether, ethoxydiglycol,polyethylene glycol-6 caprylic/capric glyceride, ethylene glycolmonobutyl ether, polyethylene glycol-8 caprylic/capric glycerides,3-methoxy-3-methyl-1-butanol, dimethyl isosorbide, and mixtures thereof.Preferably, the water dispersible component is propylene glycol.

In the present invention the non-formaldehyde-donating preservatives areselected from ammonium benzoate, ammonium propionate,benzisothiazolinone, benzoic acid, benzotriazole, benzyl alcohol,benzylparaben, 5-bromo-5-nitro-1,3-dioxane,2-bromo-2-nitropropane-1,3-diol, butyl benzoate, butylparaben, calciumbenzoate, calcium paraben, calcium propionate, calcium salicylate,calcium sorbate, chlorhexidine diacetate, chlorhexidine digluconate,chlorhexidine dihydrochloride, chioroacetamide, chlorobutanol,p-chloro-m-cresol, chlorophene, p-chlorophenol, chlorophenesin,chlorothymol, chloroxylenol, m-cresol, o-cresol, p-cresol, dehydroaceticacid, dibromopropamidine diisethionate, dimethyl oxazolidine,dithiomethylbenzamide, domiphen, ethyl ferulate, ethylparaben, ferulicacid, glyoxal, hexamidine, hexamidine diparaben, hexamidine paraben,4-hydroxybenzoic acid, hydroxymethyl dioxoazabicyclooctane, iodopropynylbutylcarbamate, isobutylparaben, isodecylparaben, isopropyl cresols,isopropylparaben, isopropyl sorbate, lauryl diethylenediaminoglycineHCl, magnesium benzoate, magnesium propionate,methyl-chloroisothiazolinone, methylparaben, octylisothiazolinone,panthenyl ethyl ether benzoate, phenethyl alcohol, phenol,phenoxyethanol, phenoxyethylparaben, phenoxyisopropanol, phenylbenzoate, phenylparaben, o-phenylphenol, polymethoxy bicyclicoxazolidine, potassium benzoate, potassium butylparaben, potassiumethylparaben, potassium methylparaben, potassium paraben, potassiumphenoxide, potassium propionate, potassium propylparaben, potassiumsorbate, propionic acid, propyl benzoate, propylparaben, quaternium-8(methyl and stearyl dimethylaminoethyl methacrylate quaternized withdimethyl sulfate), quaternium-14 (ethanaminium,N,N,N-trimethyl-2-[(2-methyl-1-oxo-2-propenyl)oxy]-, methyl sulfate,homopolymer), quaternium-15 (ethanaminium,N,N,N-trimethyl-2-[(2-methyl-1-oxo-2-propenyl)oxy]-chloride, polymerwith 2-propenamide), sodium benzoate, sodium butylparaben, sodiump-chloro-m-cresol, sodium dehydroacetate, sodium ethylparaben, sodiumformate, sodium hydroxymethane sulfonate, sodium hydroxymethylglycinate,sodium isobutylparaben, sodium isopropylparaben, sodium lauryldiethylenediaminoglycinate, sodium methylparaben, sodium paraben, sodiumphenylsulfonate, sodium phenoxide, sodium o-phenylphenate, sodiumpropionate, sodium propylparaben, sodium sorbate, sorbic acid,TEA-sorbate (triethanolamine sorbate), thianthol(2,7-dimethyl-thianthrene), triclocarban, triclosan, and undecylenoylPEG5 paraben (ester of undecylenic acid and PEG5 paraben), and mixturesthereof. Preferably, the non-formaldehyde-donating preservative isbenzoic acid.

Another object of the present invention is the use of the compositionsof the present invention in the treatment or prevention of skin and skinstructure infections in a human or an animal. Accordingly, the presentinvention provides the use of ointments and creams of the presentinvention in the treatment or prevention of skin and skin structureinfections, being non-limitative examples of such skin and skinstructure infections impetigo, foliculitis, forunculosis, acne,secondarily-infected traumatic lesions, overinfected dermatoses, andsecondarily-infected burns, and those skin and skin structure infectionscaused by methicillin-susceptible Staphylococcus aureus (MSSA),methicillin-resistant Staphylococcus aureus (MRSA) includingciprofloxacin-resistant strains, methicillin-susceptible Staphylococcusepidermidis (MSSE), methicillin-resistant Staphylococcus epidermidis(MRSE), Streptococcus pyogenes, and Group G Streptococci.

Another object of the present invention is the use of new compositionsin the treatment or prevention of sexually transmitted diseases andgenital tract infections in a human or an animal. Accordingly, thepresent invention provides the use of ointments and creams of thepresent invention in the treatment or prevention of sexually transmitteddiseases and genital tract infections, such as those caused byStreptococcus agalactiae group B, Neisseria gonorrhoeae, Chlamydiatrachomatis, Mycoplasma hominis, and Ureaplasma urealyticum.

Another object of the present invention is the use of new compositionsin the eradication of nasopharynx infections in asymptomatic nasalcarriers in a human or an animal. Accordingly, the present inventionprovides the use of ointments and creams of the present invention in theeradication of nasopharynx infections in asymptomatic nasal carriers,the infections being caused by methicillin-susceptible Staphylococcusaureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA)including ciprofloxacin-resistant strains, penicillin-resistantStreptococcus pneumoniae, Beta-lactamase positive Haemophilusinfluenzae, non-typeable strains of Haemophilus influenzae,Beta-lactamase positive Moraxella catarrhalis, Neisseria meningitides,Legionella pneumophila, Mycoplasma pneumoniae, Legionella pneumophila,and Mycobacterium tuberculosis.

The compositions of the present invention may be used by directapplication to the affected or to protect skin or genital area. Also thecompositions may be used by administration to the nasal cavity,preferably to the nasopharynx, in particular the anterior nasopharynx.

Also, the compositions of the present invention may be used in thetreatment of skin and skin structure infections, sexually transmitteddiseases and genital tract infections, and in the eradication ofnasopharynx infections in asymptomatic nasal carriers when suchinfections are resistant to usual topical antibiotics, being suchantibiotics mupirocin, fusidic acid, retapamulin, and quinolonecompounds, i.e. nadifloxacin.

Another object of the present invention is to provide novel methods totreat or prevent skin and skin structure infections in a human or ananimal in need by administering the compositions of the presentinvention.

Another object of the present invention is to provide novel methods totreat or prevent sexually transmitted diseases and genital tractinfections in a human or an animal in need by administering thecompositions of the present invention.

Another object of the present invention is to provide novel methods forthe eradication of nasopharynx infections in asymptomatic nasal carriersin a human or an animal in need by administering the compositions of thepresent invention.

The compositions according to the invention can be used effectively andsafely without clinically significant dermatological or systemic relatedadverse events, because of the skin absorption of ozenoxacin isnegligible.

Throughout the description and claims the word “comprise” and variationsof the word, such as “comprising”, are not intended to exclude othertechnical features, additives, components, or steps. Additional objects,advantages and features of the invention will become apparent to thoseskilled in the art upon examination of the description or may be learnedby practice of the invention. The following examples are provided by wayof illustration, and are not intended to be limiting of the presentinvention.

EXAMPLES Example 1 Ointment Containing 1% of Ozenoxacin

a) 100 g Composition

Ozenoxacin  1 g White soft paraffin 99 g

b) Manufacturing

White soft paraffin (99 parts) was melt to homogeneity at 70-75° C. in areactor of adequate capacity for the batch manufacturing, equipped withlow speed stirring (anchor stirring) and high speed stirring, and asystem of heating and cooling. The paraffin was cooled at 50-55° C.Ozenoxacin (1 part) was added and dispersed into paraffin by stirring.The final dispersion was cooled to 25-30° C. A homogeneous, yellow pale,oily-like base ointment was obtained. The final ointment was packed inaluminium 20 mL tubes by using an automatic system.

c) Stability

The inert nature of the soft paraffin carrier warrants that the ointmentcan be stored stably for 18 months at least.

Example 2 Cream Containing 1% of Ozenoxacin

a) 100 g Composition

Ozenoxacin 1 g (1%) PEG6 stearate and glycol stearate and PEG32 stearate20 g (20%) (Tefose ® 63) Oleoyl macrogol-6-glycerides (Labrafil ® M1944CS) 14 g (14%) 2-Octyl dodecanol (Eutanol ® G PH) 8 g (8%) Stearylalcohol (Lanette ® 18) 4 g (4%) Propylene glycol 15 g (15%) Benzoic acid0.1 g (0.1%) Purified water 37.9 g (37.9%)

b) Manufacturing

-   1. Propylene glycol (15 parts) and water (37.9 parts) were added to    a semisolid reactor of adequate capacity.-   2. The mixture was heated at 70-75° C. under slow speed stirring.-   3. A mixture of Tefose® 63 (20 parts), Labrafil® M 1944 CS (14    parts), Lanette® 18 (4 parts), and benzoic acid (0.1 parts) was melt    at 70-75° C. in a glass/aluminium beaker of adequate capacity under    low speed stirring.-   4. Final mixture of step 3 (organic phase) was added to the    semisolid reactor. The mixture was stirred at low speed with an    anchor stirring and at high speed with a high shear mixer for 5    minutes.-   5. A mixture of Eutanol® G PH (8 parts) and ozenoxacin (1 parts) was    added to a glass beaker of adequate capacity provided with an    adequate stirrer. The mixture was heated at 50-55° C. under    stirring.-   6. The suspension of step 5 was added to the emulsion of step 4 and    stirred at low speed with the anchor stirring and at high speed with    a high shear mixer for 20 minutes.-   7. The temperature of the resulting cream was let down to 25-30° C.    by stopping the heating.

Bulk homogeneity was checked.

-   8. The final cream was packed into aluminium 20 mL tubes with high    density polyethylene cap by using an automatic system.

c) Stability

Tables 1-12 summarize the stability studies performed with somedevelopment batches.

TABLE 1 Batch size: 600 g Experimental conditions: 25 ± 2° C./60% + 5%RH (1) Initial t = 6 months Appearance Conforms Conforms Activeingredient assay (%; mean) 100.91 100.61 Impurities assay (%) Unknownimpurities   n.d. (2) n.d. Total impurities n.d. n.d. Degradationproducts assay (%) n.d. n.d. Benzoic acid assay (%, mean) 102.14 105.65Particle size (D 90, microns) 4.44 3.90 Viscosity (cPs) 300406 212344 pH3.97 3.99 (1) Relative Humidity; (2) Not detected

TABLE 2 Batch size: 600 g Experimental conditions: 30 ± 2° C./65% ± 5%RH Initial t = 6 months Appearance Conforms Conforms Active ingredientassay (%; mean) 100.91 100.96 Impurities assay (%) Unknown impuritiesn.d. n.d. Total impurities n.d. n.d. Degradation products assay (%) n.d.n.d. Benzoic acid assay (%, mean) 102.14 108.76 Particle size (D 90,microns) 4.44 5.46 Viscosity (cPs) 300406 304312 pH 3.97 3.89

TABLE 3 Batch size: 600 g Experimental conditions: t = 1 t = 3 t = 6 40± 2° C./75% ± 5% RH Initial month months months Appearance ConformsConforms Conforms Conforms Active ingredient assay (%; mean) 100.9199.14 100.85 99.73 Impurities assay (%) Unknown impurities n.d. n.d.n.d. n.d. Total impurities n.d. n.d. n.d. n.d. Degradation productsassay (%) n.d. n.d. n.d. n.d. Benzoic acid assay (%, mean) 102.14 100.12102.65 108.75 Particle size (D 90, microns) 4.44 5.30 5.97 9.64Viscosity (cPs) 300406 360031 280969 398719 pH 3.97 3.91 4.10 3.89

TABLE 4 Batch size: 600 g Experimental conditions: 25 ± 2° C./60% ± 5%RH Initial t = 6 months Appearance Conforms Conforms Active ingredientassay (%; mean) 101.33 100.81 Impurities assay (%) Unknown impuritiesn.d. n.d. Total impurities n.d. n.d. Degradation products assay (%) n.d.n.d. Benzoic acid assay (%, mean) 103.85 109.0 Particle size (D 90,microns) 4.18 3.88 Viscosity (cPs) 498375 395062 pH 4.04 4.18

TABLE 5 Batch size: 600 g Experimental conditions: 30 ± 2° C./65% + 5%RH Initial t = 6 months Appearance Conforms Conforms Active ingredientassay (%; mean) 101.33 100.41 Impurities assay (%) Unknown impuritiesn.d. n.d. Total impurities n.d. n.d. Degradation products assay (%) n.d.n.d. Benzoic acid assay (%, mean) 103.85 110.03 Particle size (D 90,microns) 4.18 5.01 Viscosity (cPs) 498375 498156 pH 4.04 4.22

TABLE 6 Batch size: 600 g Experimental conditions: t = 1 t = 3 t = 6 40± 2° C./75% ± 5% RH Initial month months months Appearance ConformsConforms Conforms Conforms Active ingredient assay (%; mean) 101.3399.82 101.22 100.56 Impurities assay (%) Unknown impurities n.d. n.d.n.d. n.d. Total impurities n.d. n.d. n.d. n.d. Degradation productsassay (%) n.d. n.d. n.d. n.d. Benzoic acid assay (%, mean) 103.85 103.22107.47 111.47 Particle size (D 90, microns) 4.18 4.93 6.0 7.86 Viscosity(cPs) 498375 555500 324719 506062 pH 4.04 3.93 3.93 4.06

TABLE 7 Batch size: 8 Kg Experimental conditions: t = 3 t = 6 t = 12 25± 2° C./60% ± 5% RH Initial months months months Appearance ConformsConforms Conforms Conforms Active ingredient assay (%; mean) 102.16103.38 101.79 99.98 Impurities assay (%) Unknown impurities n.d. n.d.n.d. n.d. Total impurities n.d. n.d. n.d. n.d. Degradation productsassay (%) n.d. n.d. n.d. n.d. Benzoic acid assay (%, mean) 100.59 100.20103.71 102.54 Particle size (D 90, microns) 5.85 6.02 3.64 2.89Viscosity (cPs) 400234 209531 320094 384188 pH 4.18 4.42 4.21 4.05

TABLE 8 Batch size: 8 Kg Experimental conditions: t = 1 t = 3 t = 6 t =12 30 ± 2° C./65% ± 5% RH Initial month months months months AppearanceConforms Conforms Conforms Conforms Conforms Active ingredient assay (%;mean) 102.16 101.67 103.12 102.92 98.98 Impurities assay (%) UnknownImpurities n.d. n.d. n.d. n.d. n.d. Total Impurities n.d. n.d. n.d. n.d.n.d. Degradation products assay (%) n.d. n.d. n.d. n.d. n.d. Benzoicacid assay (%, mean) 100.59 99.70 101.38 105.21 103.26 Particle size (D90, microns) 5.85 5.5 5.03 3.68 3.79 Viscosity (cPs) 400234 350875224031 334219 338812 pH 4.18 3.95 4.05 3.95 4.04

TABLE 9 Batch size: 8 Kg Experimental conditions: t = 1 t = 3 t = 6 40 ±2° C./75% ± 5% RH Initial month months months Appearance ConformsConforms Conforms Conforms Active ingredient assay (%; mean) 102.16100.17 102.89 102.43 Impurities assay (%) Unknown impurities n.d. n.d.n.d. n.d. Total impurities n.d. n.d. n.d. n.d. Degradation productsassay (%) n.d. n.d. n.d. n.d. Benzoic acid assay (%, mean) 100.59 103.35102.28 103.21 Particle size (D 90, microns) 5.85 8.27 5.96 3.56Viscosity (cPs) 400234 439250 308437 328562 pH 4.18 3.97 3.99 3.83

TABLE 10 Batch size: 8 Kg Experimental conditions: t = 3 t = 6 t = 12 25± 2° C./60% ± 5% RH Initial months months months Appearance ConformsConforms Conforms Conforms Active ingredient assay (%; mean) 99.70102.59 100.45 99.30 Impurities assay (%) Unknown impurities n.d. n.d.n.d. n.d. Total impurities n.d. n.d. n.d. n.d. Degradation productsassay (%) n.d. n.d. n.d. n.d Benzoic acid assay (%, mean) 102.72 101.14103.71 103.06 Particle size (D 90, microns) 4.68 5.96 2.97 2.41Viscosity (cPs) 309312 264656 247500 317875 pH 4.30 4.38 3.89 4.29

TABLE 11 Batch size: 8 Kg Experimental conditions: t = 1 t = 3 t = 6 t =12 30 ± 2° C./65% ± 5% RH Initial month months months months AppearanceConforms Conforms Conforms Conforms Conforms Active ingredient assay (%;mean) 99.70 100.65 102.37 100.90 99.54 Impurities assay (%) Unknownimpurities n.d. n.d. n.d. n.d. n.d. Total impurities n.d. n.d. n.d. n.d.n.d. Degradation products assay (%) n.d. n.d. n.d. n.d. n.d. Benzoicacid assay (%, mean) 102.72 102.0 102.11 104.11 103.78 Particle size (D90, microns) 4.68 6.46 5.30 4.61 4.02 Viscosity (cPs) 309312 321250257843 304437 340750 pH 4.30 3.98 4.10 3.89 4.02

TABLE 12 Batch size: 8 Kg Experimental conditions: t = 1 t = 3 t = 6 40± 2° C./75% ± 5% RH Initial month months months Appearance ConformsConforms Conforms Conforms Active ingredient assay (%; mean) 99.70100.68 102.94 101.58 Impurities assay (%) Unknown impurities n.d. n.d.n.d. n.d. Total impurities n.d. n.d. n.d. n.d. Degradation productsassay (%) n.d. n.d. n.d. n.d. Benzoic acid assay (%, mean) 102.72 102.98103.09 104.75 Particle size (D 90, microns) 4.68 6.67 4.82 3.86Viscosity (cPs) 309312 400937 303625 310844 pH 4.30 3.97 4.02 3.86

Example 3 Antibacterial Activity of the Ointment Containing 1% ofOzenoxacin

a) Experimental Procedure

The antibacterial activity was evaluated in a model of infectionconsisting of a cutaneous wound infected by Staphylococcus aureus (ATCC6538) in mice after topical administration of the ointment of Example 1.

CD-1 male mice (45 animals) with a body weight of 22-27 g at start oftreatment were housed in standard laboratory conditions (temperature22±1° C. and relative humidity 65±10%; 12-hour light (7:00 am to 7:00pm)/12-hour dark cycle).

In order to infect the suture threads, they were submerged for 30minutes in one-night broth of S. aureus at a concentration of 10⁸CFU/mL, previously adjusted by spectrophotometry.

The threads were removed and left to dry on filter paper. Two 1 cmlengths were cut from each suture thread and each length was vortexed ina tube with 1 mL of 0.2% yeast extract. Dilutions were made of thesebroths and transferred, in duplicate and in parallel, toCystine-Lactose-Electrolyte-Deficient (CLED) agar in order to find outthe concentration on the suture threads. The remainder of the suturethread was kept in a refrigerator until it was used.

The day before the start of the experiment, the animals were shaved anddepilated with a commercial depilatory cream.

The mice were allocated at random to three groups consisting of 15animals. Groups were coded to allow a blind treatment, according toTable 13.

TABLE 13 Code # Group Treatment Concentration Form 1 A Placebo Ointment2 B Mupirocin 2% Ointment 3 C Ozenoxacin 1% Ointment

On day 1, at the start of the experiment, the animals were anaesthetizedwith isoflurane. The infection was induced with a needle threaded withsilk suture thread, previously infected with an inoculum of S. aureus ata determined concentration. A puncture was made in order that onlypierced the skin at the height of the shoulder girdle and exitedapproximately 1 cm below.

Knots were tied at each end of the thread to ensure that it did not movefrom its subcutaneous position. Then, a superficial incision was madewith a scalpel between both knots, without reaching the panniculuscarnosus.

The different treatments were applied one and eight hours after theinfection. The treatments were applied topically to the affected area.All the applications were done by massaging the infected area for noless than 30 seconds. The treatments continued for four more days andwere applied at 12-hour intervals.

The application volume was 0.1 mL/animal. The placebo was received thevehicle used in the formulation of the test item as ointment. Thetreatments will be coded before their application.

The animals were weighed and any clinical signs related to the test werenoted daily.

On day 6, approximately 16 hours after the application, all the animalswere sacrificed by cervical dislocation. An area of skin, approximately1×2 cm and which included the wound, was removed and weighed. Thissample was homogenized in 5 mL of physiological saline. This solution(0.1 mL) and three 1:10 consecutive dilutions of the initial 5-mLsolution were placed, in parallel, on plates with CLED agar+50 mM MgCl₂.In the mupirocin group, the 50 mM MgCl₂ was replaced with 2% activatedcharcoal. The two types of agar plates were used for the placebo group.

MgCl₂ was added to the CLED agar to act as a quinolone chelator in orderto inhibit the activity of the antibiotic on the plate, while activatedcharcoal (2%) was used to avoid carryover of mupirocin in the skinsamples of animals treated with 2% mupirocin ointment. The homogenizedsamples were kept in a refrigerator until the final counts had beendone, in case the counts had to be repeated.

b) Results

Values on CLED Agar+2% Activated Charcoal In the placebo group, valuesof 6.53±0.218 (mean±SEM) for Log(CFU/g skin) were obtained in thecounts. Growth was observed in all the plates.

In the 2% mupirocin ointment group, the values for Log(CFU/g skin) were4.92±0.236. Growth was observed in all plates. Statistically significantdifferences (Student t test, p<0.01) were observed between mupirocin andplacebo.

The cure rate for 2% mupirocin ointment was 24% compared to the placebogroup.

Values on CLED Agar+50 mM MgCl₂

In the placebo group, values of 6.32±0.264 (mean±SEM) for Log (CFU/gskin) were obtained in the counts. Growth was observed in all theplates.

In the 1% ozenoxacin ointment group, the values for Log (CFU/g skin)were 3.56±0.248. Growth was observed in 13 of 15 plates. Statisticallysignificant differences (Student t test, p<0.01) were observed betweenozenoxacin and placebo.

The cure rate for 1% ozenoxacin ointment was 44% compared to the placebogroup.

c) Conclusion

A 5-day administration of the ointment containing ozenoxacin 1% induceda statistically significant and higher decrease in bacterial growth inthe experimental model of an infection by Staphylococcus aureus in micethan obtained with ointment containing mupirocin 2%. No adverse localeffects were observed after application of the treatment.

Example 4 Antibacterial Activity of the Cream Containing 1% ofOzenoxacin

a) Experimental Procedure

The antibacterial activity of cream of Example 2 was evaluatedanalogously to Example 3a. The only differences concern thepharmaceutical forms, which are shown in Table 14.

TABLE 14 Code # Group Treatment Concentration Form 1 A Placebo Cream 2 BMupirocin 2% Ointment 3 C Ozenoxacin 1% Cream

b) Results

Values on CLED Agar+2% Activated Charcoal

In the placebo group, values of 6.80±0.145 (mean±SEM) for Log(CFU/gskin) were obtained in the counts. Growth was observed in all theplates.

In the 2% mupirocin ointment group, the values for Log(CFU/g skin) were5.01±0.218. Growth was observed in all plates. Statistically significantdifferences (Student t test, p<0.01) were observed between mupirocin andplacebo.

The cure rate for 2% mupirocin ointment was 26% in the observationscompared to the placebo group.

Values on CLED Agar+50 mM MgCl₂

In the placebo group, values of 6.67±0.171 (mean±SEM) for Log (CFU/gskin) were obtained in the counts. Growth was observed in all theplates.

In the 1% ozenoxacin cream group, the values for Log (CFU/g skin) were3.10±0.154. Growth was observed in 13 of 15 plates. Statisticallysignificant differences (Student t test, p<0.01) were observed betweenozenoxacin and placebo.

The cure rate for 1% ozenoxacin cream was 54% compared to the placebogroup.

c) Conclusions

A 5-day administration of the cream containing ozenoxacin 1% induced astatistically significant and higher decrease in bacterial growth in theexperimental model of infection by Staphylococcus aureus in mice thanobtained with ointment containing mupirocin 2%. No adverse local effectswere observed after application of the treatment.

Example 5 Cream Containing 2% of Ozenoxacin

100 g Composition

Ozenoxacin 2 g (2%) PEG6 stearate and glycol stearate and PEG32 stearate20 g (20%) (Tefose ® 63) Oleoyl macrogol-6-glycerides (Labrafil ® M1944CS) 14 g (14%) 2-Octyl dodecanol (Eutanol ® G PH) 8 g (8%) Stearylalcohol (Lanette ® 18) 4 g (4%) Propylene glycol 15 g (15%) Benzoic acid0.1 g (0.1%) Purified water 36.9 g (36.9%)

The manufacturing process is as for Example 2. Stability results weresimilar to those obtained for Example 2.

Example 6 Phase I Clinical Trial of Ozenoxacin 2% Cream Formulation

Objectives

The primary objective was to assess the systemic absorption followingrepeated topical applications of ozenoxacin 2% cream by analysing thepharmacokinetic parameters derived from plasma ozenoxacinconcentrations.

The secondary objectives were to assess the safety and tolerabilityafter repeated topical applications of ozenoxacin 2% cream.

Methodology

This is a multiple-dose, double blind, randomized, placebo controlledand 2-way crossover clinical trial. 20 healthy caucasian male volunteersaged 18 to 60 years were included. The dose of administration was 0.5 gozenoxacin 2% cream/90 cm². Each subject received 3 applications of 0.5g ozenoxacin 2% cream each day for 6 days and 1 single application of0.5 g ozenoxacin 2% cream on day 7, or 3 applications of placebo creamfor 6 days and 1 single application of placebo cream on day 7 at eachperiod according to a randomization code.

Blood samples for plasma ozenoxacin concentration measurements werecollected before the 1st and the 2nd applications on day 1, before the2nd applications on day 2, before the 1st and 3rd applications on day 3and day 4, before each application on day 5 and day 6, beforeapplication on day 7, and at 0.5, 1, 2, 4, 8, 12, 24, 48 and 72 hoursafter the day 7 application.

Results

After repeated topical application of 10 mg of ozenoxacin (2% cream)three times daily for seven days, all plasma ozenoxacin concentrationswere also below the limit of quantitation. Therefore, no systemicabsorption was observed.

After repeated topical applications of ozenoxacin 2% cream, preliminaryresults showed a good tolerability profile. The most commonly recordedadverse events were application site pruritus, and erythema. No seriousadverse events were reported. All adverse events were classified as mildor moderate in intensity.

It can be concluded that ozenoxacin 2% cream is well tolerated anddermal absorption is negligible.

Embodiments of the Invention:

1. A stable topical ointment composition consisting of: a) 0.2-5% byweight (based on the total weight of the composition) of ozenoxacin, andb) a suitable carrier selected from white wax, white soft paraffin, andmixtures thereof.
 2. The stable topical ointment composition accordingto claim 1, wherein the suitable carrier is white soft paraffin.
 3. Thestable topical ointment composition according to claim 1 containing 0.5to 2% by weight of ozenoxacin.
 4. The stable topical ointmentcomposition according to claim 3 containing 1% by weight of ozenoxacin.5. The stable topical ointment composition according to claim 1 for usein the treatment or prevention of skin and skin structure infections. 6.The stable topical ointment composition according to claim 1 in thetreatment of sexually transmitted diseases and genital tract infections.7. The stable topical ointment composition according to claim 1 in theeradication of nasopharynx infections in asymptomatic nasal carriers. 8.A method for the treatment or prevention of skin and skin structureinfections comprising administering the composition according to claim 1to a subject in need thereof.
 9. A method for the treatment orprevention of sexually transmitted diseases and genital tract infectionscomprising administering the composition according to claim 1 to asubject in need thereof.
 10. A method for eradication of nasopharynxinfections in asymptomatic nasal carriers comprising administering thecomposition according to claim 1 to a subject in need thereof.
 11. Astable topical ointment composition comprising: a) 0.2-5% by weight(based on the total weight of the composition) of ozenoxacin, and b) asuitable carrier selected from white wax, white soft paraffin, andmixtures thereof.
 12. The stable topical ointment composition accordingto claim 11, wherein the suitable carrier is white soft paraffin. 13.The stable topical ointment composition according to claim 11 containing0.5 to 2% by weight of ozenoxacin.
 14. The stable topical ointmentcomposition according to claim 13 containing 1% by weight of ozenoxacin.15. The stable topical ointment composition according to claim 11 foruse in the treatment or prevention of skin and skin structureinfections.
 16. The stable topical ointment composition according toclaim 11 in the treatment of sexually transmitted diseases and genitaltract infections.
 17. The stable topical ointment composition accordingto claim 11 in the eradication of nasopharynx infections in asymptomaticnasal carriers.
 18. A method for the treatment or prevention of skin andskin structure infections comprising administering the compositionaccording to claim 11 to a subject in need thereof.
 19. A method for thetreatment or prevention of sexually transmitted diseases and genitaltract infections comprising administering the composition according toclaim 11 to a subject in need thereof.
 20. A method for eradication ofnasopharynx infections in asymptomatic nasal carriers comprisingadministering the composition according to claim 11 to a subject in needthereof.